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After incubation for 4 h, 1% Triton-X 100 was added to lysate 293T cells

This less conserved domain, compared to the N-terminal domain, notably contains a well-conserved N-capping box (S/TXXD) including the S/T N-cap residue (Figure 3A) and a hydrophobic staple motif located at the N-terminal part of 6 helix

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the pI of a protein may be determined by a variety of methods including but not limited to, isoelectric focusing and various computer algorithms (see, e.g., Bjellqvist et al., 1993 , Electrophoresis 14:1023)